Tripeptidyl peptidase inhibitors

ABSTRACT

The present invention is concerned with novel compounds of formula (I) which are inhibitors of a membrane tripeptidyl peptidase responsible for the inactivation of endogenous neuropeptides such as cholecystokinis (CCKs). The invention further relates to methods for preparing such compounds, pharmaceutical compositions comprising said compounds as well as the use as a medicine of said compounds. 
                         
wherein n is an integer 0 or 1; X represents O; S; or —(CR 4 R 5 ) m — wherein m is an integer 1 or 2; R 4  and R 5  are each independently from each other hydrogen or C 1-4 alkyl; R 1  is C 1-6 alkylcarbonyl optionally substituted with hydroxy; C 1-6 alkyloxycarbonyl; aminoC 1-6 alkylcarbonyl wherein the C 1-6 alkyl group is optionally substituted with C 3-6 cycloalkyl; mono- and di(C 1-4 alkyl)aminoC 1-6 alkylcarbonyl; aminocarbonyl substituted with aryl; C 1-6 alkylcarbonyloxyC 1-6 alkylcarbonyl; C 1-6 alkyloxycarbonylaminoC 1-6 alkylcarbonyl wherein the amino group is optionally substituted with C 1-4 alkyl; an amino acid; C 1-6 alkyl substituted with amino; or arylcarbonyl; R 2  is an optionally substituted 5-membered heterocycle, or R 2  is optionally substituted benzimidazole; R 3  is a bivalent radical —CH 2 CH 2 — optionally substituted with halo or phenylmethyl; or R 3  is a bivalent radical of formula

This application is a 371 of PCT/EP01/12388, filed Oct. 24, 2001, whichclaims priority to U.S. Provisional Application 60/244,223, filed Oct.30, 2000.

The present invention is concerned with novel compounds of formula (I)which are inhibitors of a membrane tripeptidyl peptidase responsible forthe inactivation of endogenous neuropeptides such as cholecystokinins(CCKs). The invention further relates to methods for preparing suchcompounds, pharmaceutical compositions comprising said compounds as wellas the use as a medicine of said compounds.

Cholecystokinins (CCKs) are a family of hormonal and neuronal peptideswhich exert pleiotropic biological effects in the gut and brain. Theactions of CCK are mediated by CCK_(A) and CCK_(B) receptors. CCK isknown to have a physiological role in the control of food intake, whichis enhanced by CCK_(A) agonists (Smith G. P. et al., J. Ann. N.Y. Acad.Sci., 713, 236–241 (1994)), and the control of anxiety, which isdecreased by CCK_(B) antagonists (Woodruff G. et al., Rev. Pharmac., 31,469–501 (1991)).

Tripeptidyl peptidase II (TPP II) is a CCK inactivating peptidase. TPPII is found in neurons responding to cholecystokinin as well as innon-neuronal cells. TPP II is considered to be a neuropeptidaseresponsible for CCK-8 inactivation (Rose C. et al., Nature, 380,403–409, (1996)).

TPP II could be involved in CCK-8 inactivation in the gastrointestinaltract. Exogenous CCK reduces food intake and elicits other behaviouralconcomitants of satiation. Food intake is increased by systemicadministration of CCK_(A) receptor agonists (Smith G. P. et al., J. Ann.N. Y. Acad. Sci., 713, 236–241 (1994)). Endogenous CCK-controlling foodintake seems to be of neuronal rather than hormonal origin and acts uponperipheral CCK_(A) receptors on vagal afferent fibres (Smith G. P. etal., Am. J. Physiol., 249, R638–R641 (1985)).

Inhibitors of TPP II are useful tools in investigating the functions ofCCK neurons and may be useful drugs for the treatment of disorders suchas over-eating, obesity, problems with gastrointestinal motility andpsychotic syndromes.

WO-96/35805, published 14 Nov. 1996, discloses inhibitors of a membranetripeptidylpeptidase responsible for the inactivation of endogenousneuropeptides useful in treatment of gastrointestinal and mentaldisorders. WO-99/33801, published 8 Jul. 1999, disclosesCCK-inactivating tripeptidyl peptidase (TTP II) inhibiting compoundsuseful in the treatment of eating disorders, obesity, psychoticsyndromes and associated psychiatric disorders.

The compounds of the present invention differ from the cited art-knowncompounds structurally, by the nature of the R² substituent.

The present invention concerns compounds of formula (I)

a stereochemically isomeric form thereof, or a pharmaceuticallyacceptable addition salt thereof, wherein

-   -   n is an integer 0 or 1;    -   X represents O; S; or —(CR⁴R⁵)_(m)— wherein m is an integer 1 or        2; R⁴ and R⁵ are each independently from each other hydrogen or        C₁₋₄alkyl;    -   R¹ is C₁₋₆alkylcarbonyl optionally substituted with hydroxy;        C₁₋₆alkyloxycarbonyl; aminoC₁₋₆alkylcarbonyl wherein the        C₁₋₆alkyl group is optionally substituted with C₃₋₆cycloalkyl;        mono- and di(C₁₋₄alkyl)aminoC₁₋₆alkylcarbonyl; aminocarbonyl        substituted with aryl; C₁₋₆alkylcarbonyloxyC₁₋₆alkylcarbonyl;        C₁₋₆alkyloxycarbonylaminoC₁₋₆alkylcarbonyl wherein the amino        group is optionally substituted with C₁₋₄alkyl; an amino acid        residue bound via the carbonyl group; C₁₋₆alkyl substituted with        amino; or arylcarbonyl;    -   R² is a 5-membered heterocycle selected from

-   -   -   wherein m′ is an integer 1 to 2;        -   R⁶ is hydrogen or C₁₋₄alkyl;        -   R⁷ is independently from each other hydrogen; halo; amino;            hydroxy; trifluoromethyl; C₁₋₆alkyl; C₁₋₄alkyl substituted            with hydroxy, hydroxycarbonyl, C₁₋₄alkyloxycarbonyl,            aminocarbonyl, mono- or di(C₁₋₄alkyl)aminocarbonyl, amino,            or mono- or di(C₁₋₄alkyl)amino; phenyl; aminocarbonyl;            hydroxycarbonyl; C₁₋₄alkyloxycarbonyl; C₁₋₄alkylcarbonyl; or            C₁₋₄alkyloxycarbonylC₁₋₄alkylaminocarbonyl;

    -   or R² is benzimidazole, or benzimidazole substituted with one or        two substituents each independently selected from halo,        trifluoromethyl, C₁₋₄alkyl, hydroxy, hydroxycarbonyl, or        C₁₋₄alkyloxycarbonyl;

    -   R³ is a bivalent radical —CH₂CH₂— optionally substituted with        halo or phenylmethyl;

    -   or R³ is a bivalent radical of formula

-   -   -   wherein said (b-1), (b-2), or (b-3) optionally can be            substituted with one, two or three substituents each            independently selected from halo, hydroxy, C₁₋₆alkyl,            C₁₋₆alkyloxy, nitro, amino, cyano, trifluoromethyl, phenyl,            or phenyl substituted with one or two subsitutents each            independently selected from halo, hydroxy, cyano, C₁₋₆alkyl,            C₁₋₆alkyloxy, nitro, cyano, and trifluoromethyl;

    -   aryl is phenyl, or phenyl substituted with amino, nitro or        hydroxycarbonyl.

The term “amino acid residues” as used herein are the glycine, alanine,valine, leucine, isoleucine, methionine, proline, phenylalanine,tryptophan, serine, threonine, cysteine, tyrosine, asparagine,glutamine, aspartic acid, esters of aspartic acid, glutamic acid, estersof glutamic acid, lysine, arginine, and histidine amino acid radicalswhich are bound via their carbonyl group to the nitrogen atom of therest of the molecule and which can be generally represented by“R—CH(NH₂)—CO—”.

As used in the foregoing definitions halo is generic to fluoro, chloro,bromo and iodo; C₁₋₄alkyl defines straight and branched chain saturatedhydrocarbon radicals having from 1 to 4 carbon atoms such as, forexample, methyl, ethyl, propyl, butyl, 1-methylethyl, 2-methylpropyl andthe like; C₁₋₆alkyl is meant to include C₁₋₄alkyl and the higherhomologues thereof having 5 or 6 carbon atoms, such as, for example,2-methylbutyl, pentyl, hexyl and the like; C₃₋₆cycloalkyl is generic tocyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl; C₃₋₆alkenyl definesstraight and branched chain unsaturated hydrocarbon radicals having from3 to 6 carbon atoms, such as propenyl, butenyl, pentenyl or hexenyl;C₁₋₂alkanediyl defines methylene or 1,2-ethanediyl; C₁₋₅alkanediyldefines bivalent straight or branched chain hydrocarbon radicalscontaining from 1 to 5 carbon atoms such as, for example, methylene,1,2-ethanediyl, 1,3-propanediyl, 1,4-butanediyl, 1,5-pentanediyl, andthe branched isomers thereof; C₁₋₆alkanediyl includes C₁₋₅alkanediyl andthe higher homologues thereof having 6 carbon atoms such as, forexample, 1,6-hexanediyl and the like. The term “CO” refers to a carbonylgroup.

The term “stereochemically isomeric forms” as used hereinbefore definesall the possible isomeric forms which the compounds of formula (I) maypossess. Unless otherwise mentioned or indicated, the chemicaldesignation of compounds denotes the mixture of all possiblestereochemically isomeric forms, said mixtures containing alldiastereomers and enantiomers of the basic molecular structure. More inparticular, stereogenic centers may have the R- or S-configuration;substituents on bivalent cyclic (partially) saturated radicals may haveeither the cis- or trans-configuration. Compounds encompassing doublebonds can have an E or Z-stereochemistry at said double bond.Stereochemically isomeric forms of the compounds of formula (I) areobviously intended to be embraced within the scope of this invention.

The pharmaceutically acceptable addition salts as mentioned hereinaboveinclude pharmaceutically acceptable acid addition salts and are meant tocomprise the therapeutically active non-toxic acid addition salt formswhich the compounds of formula (I) are able to form. Thepharmaceutically acceptable acid addition salts can conveniently beobtained by treating the base form with such appropriate acid.Appropriate acids comprise, for example, inorganic acids such ashydrohalic acids, e.g. hydrochloric or hydrobromic acid, sulfuric,nitric, phosphoric and the like acids; or organic acids such as, forexample, acetic, propanoic, hydroxyacetic, lactic, pyruvic, oxalic (i.e.ethanedioic), malonic, succinic (i.e. butanedioic acid), maleic,fumaric, malic, tartaric, citric, methanesulfonic, ethanesulfonic,benzenesulfonic, p-toluenesulfonic, cyclamic, salicylic,p-aminosalicylic, pamoic and the like acids.

Where the compounds of the invention carry an acidic moiety, suitablepharmaceutically acceptable base addition salts are possible whichinclude alkali metal salts, e.g., sodium or potassium salts; alkalineearth metal salts, e.g., calcium or magnesium salts; and base additionsalts formed with suitable organic ligands, e.g., primary, secondary,tertiary or quaternary ammonium salts, such as morpholinyl,tert-butylamino, and the like.

Conversely said salt forms can be converted by treatment with anappropriate base into the free base form.

The term addition salt as used hereinabove also comprises the solvateswhich the compounds of formula (I) as well as the salts thereof, areable to form. Such solvates are for example hydrates, alcoholates andthe like.

Interesting compounds are those compounds of formula (I) wherein one ormore of the following restrictions apply:

-   -   a) n is 0;    -   b) R³ is a radical of formula (b-1) optionally substituted with        halo or methoxy;    -   c) X represents —CH₂— or —CH₂CH₂—;    -   d) R² is a radical of formula (a-2) wherein R⁶ is hydrogen;    -   e) R² is a radical of formula (a-2), (a-4), (a-6), or (a-7);    -   f) R² is benzimidazole optionally substituted with methyl,        hydroxy, halo, trifluoromethyl, methyloxycarbonyl, or        hydroxycarbonyl;    -   g) R¹ is C₁₋₆alkylcarbonyl, aminoC₁₋₆alkylcarbonyl or an amino        acid.

Particular compounds are those compounds of formula (I) wherein n is 0and R³ is a radical of formula (b-1) optionally substituted with halo ormethoxy.

Preferred compounds are those compounds of formula (I) wherein n is 0,R³ is a radical of formula (b-1) optionally substituted with halo ormethoxy, and X represents —CH₂—.

Other preferred compounds are those compounds of formula (I) wherein nis 0, R³ is a radical of formula (b-1) optionally substituted with haloor methoxy, and X represents —CH₂CH₂—.

Still other preferred compounds are those compounds of formula (I)wherein R¹ is C₁₋₆alkylcarbonyl, aminoC₁₋₆alkylcarbonyl or an aminoacid.

Compounds of formula (I-a), defined as compounds of formula (I) whereinR^(1a) represents all R¹ substituents other than C₁₋₄alkyl substitutedwith amino, can be prepared by reacting an intermediate of formula (II)with an intermediate of formula (III) in the presence of4-methyl-morpholine, in a reaction-inert solvent such as, e.g.dichloromethane of chloroform. Stirring may enhance the rate of thereaction. The reaction may conveniently be carried out at a temperatureranging between room temperature and the reflux temperature of thereaction mixture and, if desired, the reaction may be carried out in anautoclave at an increased pressure. Optionally said reaction is followedby an acid hydrolysis step to remove acid labile protecting groups, suchas a tert-butyloxycarbonyl.

Alternatively, compounds of formula (I-a) can also be prepared byreacting an intermediate of formula (II) with an intermediate of formula(IV) in the presence of an appropriate activating agent, such as e.g.isobutyl chloroformate, in a reaction-inert solvent such as, e.g.dichloromethane, in the presence of a suitable base such as, e.g.triethylamine. Optionally said reaction is followed by an acidhydrolysis step to remove acid labile protecting groups, such as atert-butyloxycarbonyl.

Compounds of formula (I-b), defined as compounds of formula (I) whereinR¹ represents C₁₋₆alkyl substituted with amino, can conveniently beprepared by submitting the corresponding starting compounds (I-b′)wherein R¹ represents aminoC₁₋₅alkylcarbonyl to an appropriate reductionreaction. Appropriate reduction reactions can be e.g. treatment withborane-tetahydrofuran complex.

Compounds of formula (I-c), defined as compounds of formula (I) whereinR² represents a radical.(a-2) wherein R⁶ is hydrogen and R⁷ is locatedat the 3-position of the imidazole moiety, can be prepared by reactingan intermediate of formula (V) with an intermediate of formula (VI) inthe presents of potassium acetate in a suitable solvent such asmethanol.

The compounds of formula (I) may further be prepared by convertingcompounds of formula (I) into each other according to art-known grouptransformation reactions.

The starting materials and some of the intermediates, such as e.g.intermediates of formula (III), (IV) and (VI), are known compounds andare commercially available or may be prepared according to conventionalreaction procedures generally known in the art.

Compounds of formula (I) and some of the intermediates may have one ormore stereogenic centers in their structure, present in a R or a Sconfiguration, such as, e.g. the carbon atom bearing the R² substituent.

Following CAS nomenclature conventions, when two stereogenic centers ofknown absolute configuration are present in a molecule, an R or Sdescriptor is assigned (based on Cahn-Ingold-Prelog sequence rule) tothe lowest-numbered chiral center, the reference center. Theconfiguration of the second stereogenic center is indicated usingrelative descriptors [R*,R*] or [R*,S*], where R* is always specified asthe reference center and [R*,R*] indicates centers with the samechirality and [R*,S*] indicates centers of unlike chirality. Forexample, if the lowest-numbered chiral center in the molecule has an Sconfiguration and the second center is R, the stereo descriptor would bespecified as S-[R*,S*].

The compounds of formula (I) as prepared in the hereinabove describedprocesses may be synthesized in the form of racemic mixtures ofenantiomers which can be separated from one another following art-knownresolution procedures. The racemic compounds of formula (I) may beconverted into the corresponding diastereomeric salt forms by reactionwith a suitable chiral acid. Said diastereomeric salt forms aresubsequently separated, for example, by selective or fractionalcrystallization and the enantiomers are liberated therefrom by alkali.An alternative manner of separating the enantiomeric forms of thecompounds of formula (I) involves liquid chromatography using a chiralstationary phase. Said pure stereochemically isomeric forms may also bederived from the corresponding pure stereochemically isomeric forms ofthe appropriate starting materials, provided that the reaction occursstereospecifically. Preferably if a specific stereoisomer is desired,said compound will be synthesized by stereospecific methods ofpreparation. These methods will advantageously employ enantiomericallypure starting materials.

The compounds of formula (I), the pharmaceutically acceptable salts andstereoisomeric forms thereof are inhibitors of a membrane tripeptidylpeptidase responsible for the inactivation of endogenous neuropeptidessuch as cholecystokinis (CCKs) as evidenced in pharmacological exampleC-1.

In view of their TPP II inhibiting properties the compounds of thepresent invention are useful in treatment of conditions or disordersassociated with TPP II activity such as, e.g. eating disorders, obesity,psychotic syndromes and associated psychiatric disorders.

In view of the utility of the compounds of formula (I), it follows thatthe present invention also provides a method of treating warm-bloodedanimals, including humans, (generally called herein patients) sufferingfrom eating disorders, obesity, psychotic syndromes and associatedpsychiatric disorders Consequently a method of treatment is provided forinhibiting the activity of TPP II and/or relieving patients sufferingfrom conditions, such as, for example, eating disorders, obesity,psychotic syndromes and associated psychiatric disorders.

Hence, the use of a compound of formula (I) as medicine is providedacting as an inhibitor of the CCK-inactivating peptidase tripeptidylpeptidase (TPP II) and/or for the treatment of eating disorders,especially obesity and/or for the treatment of psychotic syndromes andassociated psychiatric disorders, which comprises a therepauticallyeffective amount of a compound of formula (I). Also provided is the useof a compound of formula (I) for the manufacture of a medicine forinhibiting the activity of TPP II and/or treating eating disorders,obesity, psychotic syndromes and associated psychiatric disorders. Bothprophylactic and therapeutic treatment are envisaged.

It is believed that some of the compounds of the present invention, inparticular compounds (153) to (181), may also have opioid activity suchas delta-opioid (δ), mu-opioid (μ) and/or kappa-opioid (κ) activity.Opioid activity can be measured using the assays as described inpharmacological examples C.2 and C.3.

To prepare the pharmaceutical compositions of this invention, aneffective amount of the particular compound, in base or acid additionsalt form, as the active ingredient is combined in intimate admixturewith a pharmaceutically acceptable carrier, which carrier may take awide variety of forms depending on the form of preparation desired foradministration. These pharmaceutical compositions are desirably inunitary dosage form suitable, preferably, for administration orally,rectally or by parenteral injection. For example, in preparing thecompositions in oral dosage form, any of the usual pharmaceutical mediamay be employed, such as, for example, water, glycols, oils, alcoholsand the like in the case of oral liquid preparations such assuspensions, syrups, elixirs and solutions; or solid carriers such asstarches, sugars, kaolin, lubricants, binders, disintegrating agents andthe like in the case of powders, pills, capsules and tablets. Because oftheir ease in administration, tablets and capsules represent the mostadvantageous oral dosage unit form, in which case solid pharmaceuticalcarriers are obviously employed. For parenteral compositions, thecarrier will usually comprise sterile water, at least in large part,though other ingredients, for example, to aid solubility, may beincluded. Injectable solutions, for example, may be prepared in whichthe carrier comprises saline solution, glucose solution or a mixture ofsaline and glucose solution. Injectable suspensions may also be preparedin which case appropriate liquid carriers, suspending agents and thelike may be employed. In the compositions suitable for percutaneousadministration, the carrier optionally comprises a penetration enhancingagent and/or a suitable wetting agent, optionally combined with suitableadditives of any nature in minor proportions, which additives do notcause a significant deleterious effect to the skin. Said additives mayfacilitate the administration to the skin and/or may be helpful forpreparing the desired compositions. These compositions may beadministered in various ways, e.g., as a transdermal patch, as aspot-on, as an ointment. Acid addition salts of (I) due to theirincreased water solubility over the corresponding base form, areobviously more suitable in the preparation of aqueous compositions.

It is especially advantageous to formulate the aforementionedpharmaceutical compositions in dosage unit form for ease ofadministration and uniformity of dosage. Dosage unit form as used in thespecification and claims herein refers to physically discrete unitssuitable as unitary dosages, each unit containing a predeterminedquantity of active ingredient calculated to produce the desiredtherapeutic effect in association with the required pharmaceuticalcarrier. Examples of such dosage unit forms are tablets (includingscored or coated tablets), capsules, pills, powder packets, wafers,injectable solutions or suspensions, teaspoonfuls, tablespoonfuls andthe like, and segregated multiples thereof.

For oral administration, the pharmaceutical compositions may take theform of solid dose forms, for example, tablets (both swallowable-onlyand chewable forms), capsules or gelcaps, prepared by conventional meanswith pharmaceutically acceptable excipients such as binding agents (e.g.pregelatinised maize starch, polyvinylpyrrolidone or hydroxypropylmethylcellulose); fillers (e.g. lactose, microcrystalline cellulose orcalcium phosphate); lubricants e.g. magnesium stearate, talc or silica);disintegrants (e.g. potato starch or sodium starch glycollate); orwetting agents (e.g. sodium lauryl sulphate). The tablets may be coatedby methods well known in the art.

Liquid preparations for oral administration may take the form of, forexample, solutions, syrups or suspensions, or they may be presented as adry product for constitution with water or other suitable vehicle beforeuse. Such liquid preparations may be prepared by conventional means,optionally with pharmaceutically acceptable additives such as suspendingagents (e.g. sorbitol syrup, methylcellulose, hydroxy-propylmethylcellulose or hydrogenated edible fats); emulsifying agents (e.g.lecithin or acacia); non-aqueous vehicles (e.g. almond oil, oily estersor ethyl alcohol); and preservatives (e.g. methyl or propylp-hydroxybenzoates or sorbic acid).

Pharmaceutically acceptable sweeteners comprise preferably at least oneintense sweetener such as saccharin, sodium or calcium saccharin,aspartame, acesulfame potassium, sodium cyclamate, alitame, adihydrochalcone sweetener, monellin, stevioside or sucralose(4,1′,6′-trichloro4,1′,6′-trideoxygalactosucrose), preferably saccharin,sodium or calcium saccharin, and optionally a bulk sweetener such assorbitol, mannitol, fructose, sucrose, maltose, isomalt, glucose,hydrogenated glucose syrup, xylitol, caramel or honey.

Intense sweeteners are conveniently employed in low concentrations. Forexample, in the case of sodium saccharin, the concentration may rangefrom 0.04% to 0.1% (w/v) based on the total volume of the finalformulation, and preferably is about 0.06% in the low-dosageformulations and about 0.08% in the high-dosage ones. The bulk sweetenercan effectively be used in larger quantities ranging from about 10% toabout 35%, preferably from about 10% to 15% (w/v).

The pharmaceutically acceptable flavours which can mask the bittertasting ingredients in the low-dosage formulations are preferably fruitflavours such as cherry, raspberry, black currant or strawberry flavour.A combination of two flavours may yield very good results. In thehigh-dosage formulations stronger flavours may be required such asCaramel Chocolate flavour, Mint Cool flavour, Fantasy flavour and thelike pharmaceutically acceptable strong flavours. Each flavour may bepresent in the final composition in a concentration ranging from 0.05%to 1% (w/v). Combinations of said strong flavours are advantageouslyused. Preferably a flavour is used that does not undergo any change orloss of taste and colour under the acidic conditions of the formulation.

The compounds of the invention may also be formulated as depotpreparations. Such long acting formulations may be administered byimplantation (for example subcutaneously or intramuscularly) or byintramuscular injection. Thus, for example, the compounds may beformulated with suitable polymeric or hydrophobic materials (for exampleas an emulsion in an acceptable oil) or ion exchange resins, or assparingly soluble derivatives, for example as a sparingly soluble salt.

The compounds of the invention may be formulated for parenteraladministration by injection, conveniently intravenous, intramuscular orsubcutaneous injection, for example by bolus injection or continuousintravenous infusion. Formulations for injection may be presented inunit dosage form e.g. in ampoules or in multidose containers, with anadded preservative. The compositions may take such forms as suspensions,solutions or emulsions in oily or aqueous vehicles, and may containformulatory agents such as isotonizing, suspending, stabilising and/ordispersing agents. Alternatively, the active ingredient may be in powderform for constitution with a suitable vehicle, e.g. sterile pyrogen-freewater before use.

The compounds of the invention may also be formulated in rectalcompositions such as suppositories or retention enemas, e.g. containingconventional suppository bases such as cocoa butter or other glycerides.

For intranasal administration the compounds of the invention may beused, for example, as a liquid spray, as a powder or in the form ofdrops.

Experimental Part

In the procedures described hereinafter the following abbreviations wereused: “ACN” stands for acetonitrile; “THF”, which stands fortetrahydrofuran; “DCM” stands for dichloromethane; and “MIK” stands formethyl isobutyl ketone.

For some chemicals the chemical formula was used, e.g. CH₂Cl₂ fordichloromethane, CH₃OH for methanol, NH3 for ammonia, HCl forhydrochloric acid, NaOH for sodium hydroxide, NaHCO₃ for sodium hydrogencarbonate, and Na₂CO₃ sodium carbonate.

In those cases the stereochemically isomeric form which was firstisolated is designated as “A” and the second as “B”, without furtherreference to the actual stereochemical configuration.

Preparative liquid chromatography was performed on a semi-preparativeHPLC unit using a YMC ODS-A column (30×100 mm, 5 micron, temperature:ambient, flow rate: 35 mL/min, mobile phase: a) 10/90 acetonitrile/waterwith 0.1% trifluoroacetic acid, b) 90/10 acetonitrile/water with 0.1%trifluoroacetic acid, gradient: linear gradient from A to B over 9minutes, UV detection at 254 nm.

A. Preparation of the Intermediates

EXAMPLE A.1

a) 2,3-Dihydroxy-1H-indole-2-carboxamide (0.030 mol) was suspended intrichloromethane (400 ml). The mixture was cooled to 0° C. Triethylamine(0.045 mol) was added. Acetyl chloride (0.045 mol) was added over 2minutes. After 30 minutes, TLC showed the reaction was incomplete. Whilethe flask was still cool, more Triethylamine (6.26 ml) was added,followed 15 minutes later with more acetyl chloride (3.21 ml). TLCshowed the reaction was still incomplete. The reaction was continued toallow to stir, cooled to 0° C., and more triethylamine (6.26 ml) wasadded. Over 2 minutes, more acetyl chloride (3.21 ml) was added neat.TLC showed 80% completion after 60 minutes, and no progress after 30more minutes. A third portion of acetyl chloride and triethylamine wasadded. After an additional 15 minutes, ice cold water (200 ml) wasadded. The mixture was stirred for 10 minutes, filtered, and rinsed withwater (3×100 ml) and trichloromethane (2×75 ml). The sample was allowedto dry overnight, yielding 4.71 g of(S)-1-acetyl-2,3-dihydro-1H-indole-2-carboxamide (intermediate 1,mp. >260° C.).

b) Intermediate (1) (0.02022 mol) was suspended in DCM (175 ml). Themixture was cooled to 0° C. Triethylamine (0.06066 mol) was added neat.Trichloroacetyl chloride (0.03033 mol) in DCM (20 ml) was added dropwiseover 20 minutes. After 2 hours, ice water (200 ml) was added, the phasesseparated and the organic phase reextracted with 3 N HCl and then with asaturated aqueous NaHCO₃ solution. The organic phase was dried,filtered, and stripped to leave 4.61 g brown solid. The solid wastriturated with ice cold diethylether (30 ml), filtered, and rinsed withice cold diethylether (twice), yielding 3.12 g (83%) of(S)-1-acetyl-2,3-dihydro-1H-indole-2-carbonitrile (intermediate 2, mp.134–135° C.).

c) Intermediate (2) (0.0151 mol) was suspended in diethylether (200 ml).Ethanol (0.0214 mol) was added, and the mixture was cooled to 0° C. HCl(gas) was bubbled in for 45 minutes. The mixture was removed from theice bath and stirred. After 20 minutes, a residue collected on the wallsides. The walls were scratched, and a white solid precipitated out.After 1 hour the sample was filtered, rinsed with diethylether, airdried quickly, yielding 3.99 g of (S)-ethyl1-acetyl-2,3-dihydro-1H-indole-2-carboximidate monohydrochloride(intermediate 3).

In analogy, ethyl 1-acetyl-2,3-dihydro-1H-indole-2-carboximidatemonohydrochloride (intermediate 6) was prepared starting from1-acetyl-2,3-dihydro-1H-indole-2-carbonitrile.

EXAMPLE A.2

a) 5-chloro-2,3-dihydroxy-1H-indole-2-carboxylic acid, methyl ester(0.00761 mole) was dissolved in methanol (25 ml) and cooled to 0° C. NH₃was bubbled in for 10 minutes. The flask was stoppered and allowed towarm to room temperature. The mixture was stirred overnight. TLC showedthe reaction was mostly complete. The sample was concentrated to ±⅓volume, cooled, and filtered, rinsing resulting solid with ice coldmethanol (2 ml) and then dried ion the air, yielding 0.74 g of5-chloro-2,3-dihydro-1H-indole-2-carboxamide (intermediate 7, mp.151–152° C.).

b) Triethylamine (0.02080 mole) was added to intermediate (7) (0.09632mole) dissolved in trichloromethane (700 ml). The mixture was cooled to5° C. Acetyl chloride (0.2480 mole) was added over 2 minutes withstirring. After 5 minutes, a precipitate formed. The ice bath wasremoved, and the container allowed to sit for 15 minutes. Ice water (250ml) was added, and the mixture was stirred for 10 minutes. The samplewas filtered, rinsed with water and trichloromethane. The solid wassuspended in water (200 ml), and swirled for 10 minutes.Trichloromethane (200 ml) was added and the mixture was stirred, thenfiltered and rinsed with water and trichloromethane, and then dried tothe air overnight, yielding 19.71 g of1-acetyl-5-chloro-2,3-dihydro-1H-indole-2-carboxamide (intermediate 8).

c) Triethylamine (0.41291 mole) was added to intermediate (8) (0.08258mole) suspended in dichloromethane (500 ml) at 0° C. Trichloroacetylchloride (0.20645 mole) was added over 10 minutes. When the reactionappeared sluggish, an additional portion of triethylamine (20 ml) andthen more trichloroacetyl chloride (7.6 ml) were added, and the mixturewas stirred for 2 hours at low temperature. The ice bath was removed,and the mixture was allowed to sit for 2 hours. This resulted in adarker colored reaction, which was re-cooled to 0° C. Ice cold water(150 ml) was slowly added, and the mixture was stirred for 5 minutes.The layers were separated, and the organic phase was washed (ice cold 3NHCl, saturated NaHCO₃), dried, filtered, and concentrated. The residuewas triturated in ice cold diethylether (40 ml). Filtration, rinsingwith ice cold diethylether (10 ml), yielding 15.13 g of1-acetyl-5-chloro-2,3-dihydro-1H-indole-2-carbonitrile (intermediate 9,mp. 140–142° C.).

d) HCl (as a 2 M solution) was added slowly to intermediate (9) untilgas evolution was noted. Then stopped adding the prepared HCl (2N indiethylether), and suspended HCl in diethylether (150 ml) and thenethanol (0.042 mole) was added. The mixture was cooled to 0° C. and HCl(gas) was added over an hour, with an oil precipitating out. Thereaction was diluted to 1 l with diethylether. More oil precipitates,and no solid formed after sitting for 1 hour. The diethylether wasdecanted off. The residue was diluted (diethylether, 500 ml). The solidbegins to form, and the mixture was stirred for 2 hours. The sample wasfiltered, rinsing with diethylether. The sample was placed under vacuum,yielding 6.41 g of ethyl1-acetyl-5-chloro-2,3-dihydro-1H-indole-2-carboximidatemonohydrochloride (intermediate 10).

EXAMPLE A.3

a) Bis (1,1-dimethylethyl)ester dicarbonic acid (0.07615 mol) in DCM (50ml) was added over 5 minutes to 2,3-dihydro-1H-indole-2-methanol(0.07615 mol) in DCM (150 ml) at 0° C. The mixture was allowed to warmto room temperature and stirred overnight. The mixture was concentratedunder reduced pressure and submitted to a Kogel Rohr distillation,yielding 11.98 g of 1,1-dimethylethyl2,3-dihydro-2(hydroxymethyl)-1H-indole-1-carboxylate (intermediate 11).

b) Dess-Martin Reagent (0.011 mol) was added neat over 1 minutes tointermediate (11) (0.010 mol) dissolved in DCM (35 ml). After 15minutes, the ice bath was removed, and the mixture was allowed to warmto room temperature. More Dess-Martin Reagent (0.33 g) was added, andthe mixture was stirred for 30 minutes more. The mixture was re-cooledto 0° C. and treated slowly with a partial suspension/solution ofNa₂S₂O₃ (25 g) which had attempted to dissolve in a saturated aqueousNaHCO₃ (100 ml) solution. After 10 minutes, the mixture was removed fromice, and the layers were separated. More DCM was added, and the mixturewas filtered. The organic was separated from the filtrate, and thecombined organic phases were dried, filtered, concentrated and purifiedthrough flash column chromatography (eluent: 10% ethyl acetate: hexane,dissolving the sample in 3:1 ethyl acetate: hexane (5 ml)), yielding1,1-dimethylethyl 2-formyl-2,3-dihydro-1H-indole-1-carboxylate(intermediate 12, mp. 85–87° C.).

EXAMPLE A.4

A solution of 1-acetyl-2,3-dihydro-1H-indole-2-carbonitrile (0.00988mol) and Triethylamine (0.0197 mol) in pyridine (50 ml) was treated withhydrogen sulfide (gas) at room temperature via a bubbler for 2 hours andthe resultant saturated reaction mixture was closed and allowed to setfor 16 hours. The reaction mixture was poured into 200 ml of an icewater slurry. A voluminous precipitate formed. The mixture was recooledin an ice bath and the precipitate was collected by suction filtration,washed with cold water, and air dried, yielding 1.62 g of1-acetyl-2,3-dihydro-1H-indole-2carbothioamide (intermediate 13, mp.194–195° C.).

EXAMPLE A.5

1-acetyl-2,3-dihydro-1H-indole-2-carbonitrile (0.0132 mole) was treatedwith water (54 ml) and the resulting suspension was treated sequentiallywith Na₂CO₃ (0.00726 mole) and NH₂OH.HCl 0.0145 mole). The mixture wastreated with ethanol (26 ml) and heated to 80–90° C. Upon achievingreaction temperature, the mixture was still a suspension. Added another26 ml of ethanol which afforded a clear solution. The reaction washeated for 2.5 hours and cooled to room temperature with stirring. Avoluminous precipitate formed which was collected by suction filtration,washed with cold distilled water, and air dried, yielding 2.23 g of1-acetyl-2,3-dihydro-N′-hydroxy-1H-indole-2-carboximidamide(intermediate 14, mp. 204–205° C.).

EXAMPLE A.6

1-acetyl-2-(4-ethyl-1H-imidazol-2-yl)-2,3-dihydro-1H-indole (0.0035 mol)and HCl, 6N (50 ml) were combined under nitrogen atmosphere. Thereaction mixture was heated immediately and the heating was continuedfor 3.5 hours. The mixture was allowed to cool to room temperature, thenextracted with diethylether (2×75 ml), cooled to 0° C., alkalized (withcooled 3 N NaOH), then extracted with chloroform (3×60 ml). The combinedorganic layers were dried, filtered and the solvent was evaporated,yielding 0.79 g of 2-(4-ethyl-1H-imidazol-2-yl)-2,3-dihydro-1H-indole(intermediate 15).

EXAMPLE A.7

a) To a suspension of 5-fluoro-1H-indole-2-carboxylic acid, ethyl ester(0.121 mole) in methanol (600 ml) was added Mg (0.36 mole). The mixturewas in a 3-neck round bottom flask under argon at room temperature. Thetemperature of the reaction was monitored closely. After about 10minutes, the mixture began to bubble, slowly at first and then morevigorously. The reaction temperature was maintained between 15 and 25°C. with intermittent applications of an ice bath. After 30 minutes, thebubbling had slowed. The mixture was allowed to stir at room temperaturefor three days. The mixture was partitioned between 600 ml of chloroformand 500 ml of saturated NH₄Cl solution. The organic layer was dried overMgSO₄ and concentrated to a brown oil. The oil was dissolved in etherand extracted with 3N HCl. The aqueous layer was washed with ether,basified with 3N NaOH, and extracted with chloroform. The extract wasdried over MgSO₄ and concentrated, yielding 13.91 g of methyl5-fluoro-2,3-dihydro-1H-indole-2-carboxylate (intermediate 16).

b) To 2M NH₃ in methanol (0.6 mol), cooled in an ice bath under Ar, wasadded intermediate (16) (0.0574 mol) dissolved in methanol (150 ml). Themixture was allowed to warm to room temperature and stir under argon for6 hours. The reaction was concentrated to 150 ml and filtered. The solidwas rinsed with a small amount of cold methanol and allowed to dry,yielding 2.33 g of 5-fluoro-2,3-dihydro-1H-indole-2-carboxamide(intermediate 17, mp. 197–199° C.).

c) To a mixture of intermediate (17) (0.0094 mole) in DCM (30 ml),cooled in an ice bath under argon, was added triethylamine (0.031 mole)followed by acetyl chloride (0.031 mole). The resulting mixture wasallowed to return to room temperature. After stirring for 6 hours, themixture was cooled in an ice bath and 50 ml of water was added. Themixture was allowed to stir about 20 minutes, was filtered and the solidwas allowed to dry to obtain 1.58 g of1-acetyl-5-fluoro-2,3-dihydro-1H-indole-2-carboxamide (intermediate 18,mp. 232–235° C.).

d) To a suspension of intermediate (18) (0.0076 mole) in DCM (30 ml),cooled in an ice bath under argon, was added triethylamine (0.0228 mole)followed by trichloroacetyl chloride (0.0115 mole). The mixture wasallowed to warm to room temperature and stir for 2 hours. The mixturewas washed with water, 2N HCl, and saturated NaHCO₃. The organic layerwas dried and concentrated. The concentrate was triturated in ether andpurified on silica gel column, eluting with 50% ethyl acetate in hexane.The desired fractions were combined and concentrated. The residue wastriturated in ether and the solid collected by filtration and allowed todry, yielding 0.30 g of1-acetyl-5-fluoro-2,3-dihydro-1H-indole-2-carbonitrile (intermediate 19,mp. 93–95° C.)

e) A solution of intermediate (19) (0.004 mole) and HCl/diethylether (60mL) was cooled in an ice bath under argon. Ethanol (0.0075 mole) wasadded. HCl was bubbled into the solution for 50 minutes until themixture became homogeneous. The mixture was allowed to slowly warm toroom temperature and stir for 4 hours. The ether was decanted off anddissolved in methanol. The methanol solution was concentrated in vacuumand the residue was used as is for the next step, yielding ethyl1-acetyl-5-fluoro-2,3-dihydro-1H-indole-2-carboximidatemonohydrochloride (intermediate 20).

EXAMPLE A.8

a) 2,3-dihydro-5-methoxy-1H-indole-2-carboxylic acid methyl ester (0.084mole) and 2M NH₃ in methanol (500 ml) were combined and stirred at roomtemperature under argon over the weekend. The solution was concentratedto 100 ml, cooled in an ice bath, and filtered. The solid was rinsedwith a small amount of cold methanol and dried. The residue wastriturated in methanol/ACN and filtered, yielding 4.56 g of2,3-dihydro-5-methoxy-1H-indole-2-carboxamide (intermediate 21, mp.228–229° C.).

b) Triethylamine (0.0106 mole) then acetyl chloride (0.0106 mole) wereadded to a solution of intermediate (21) (0.0032 mole) in DCM (40 ml)cooled in an ice bath under argon. The mixture was allowed to slowlywarm to room temperature and stir overnight. The mixture was cooled inan ice bath and ice cold water (30 ml) was added. After stirring for 10minutes, the mixture was filtered, and the solid was allowed to dryovernight. The residue was suspended in 50 ml water. The suspension wasallowed to stir for 30 minutes, filtered, and dried overnight, yielding0.40 g of 1-acetyl-2,3-dihydro-5-methoxy-1H-indole-2-carboxamide(intermediate 22, mp. 196–197° C.).

c) To a suspension of intermediate (22) (0.022 mole) in DCM (150 ml),cooled in an ice bath under argon, was added triethylamine (0.066 mole)then trichloroacetyl chloride (0.033 mole). The mixture was allowed toslowly warmed to room temperature overnight. The mixture was washed withwater, 2N HCl, and saturated NaHCO₃. The organic phase was dried,concentrated and triturated in ether and the solid collected, yielding1-acetyl-2,3-dihydro-5-methoxy-1H-indole-2-carbonitrile (intermediate23, mp. 108–110° C.).

d) To a solution of intermediate (23) (0.0154 mole) and ethanol (0.0231mole) in 1M HCl/diethylether (200 ml), cooled in an ice bath was bubbledHCl (gas) for 60 minutes. The ice bath was maintained for 45 minutes,and the mixture was concentrated at room temperature under vacuum to 200ml of an oily precipitate. The residue was triturated to a brown solidthat became an oil after decanting off the diethylether. The residue waswashed with diethylether twice, dissolved in methanol, and used withoutfurther purification for further synthesis, yielding ethyl1-acetyl-2,3-dihydro-5-methoxy-1H-indole-2-carboximidatemonohydrochloride (intermediate 24).

EXAMPLE A.9

(S)-2-(Tert-butoxycarbonylamino)butyric acid (0.010 mol) dissolved inDCM (25 ml) was placed in a cooling bath at −10° C. Pyridine (0.010 mol)was added, followed by 2,4,6-trifluoro-1,3,5-triazine (0.0345 mol). Themixture was stirred under nitrogen. After one hour, ice cold water (75ml) was added. More DCM (45 ml) was added, and the mixture was shaken.The organic phase was separated, washed with ice cold water again (100ml), then the organic phase dried, filtered, and concentrated to yield2.29 g of (S)-1,1-dimethylethyl [1-(fluorocarbonyl)propyl]-carbamate(intermediate 25).

EXAMPLE A.10

Compound (8) (0.00170 mol) was dissolved in HCl, 6N (20 ml), andimmediately warmed in an oil bath at 100° C. under nitrogen for 200minutes. The heat was turned off, and the sample was cooled to 0° C. 3 NNaOH (35 ml) was slowly added. Basification was completed with saturatedNaHCO₃. The sample was extracted with chloroform. The combined organicphases were dried, filtered, and the resulting solution was used withoutfurther purification in further synthesis, yielding(S)-2,3-dihydro-2-(4-propyl-1H-imidazol-2-yl)-1H-indole (intermediate5).

EXAMPLE A.11

A mixture of intermediate (13) (0.00844 mol) in ethanol (180 ml) wastreated with 1-bromo-2-butanone (0.0085 mol) in one portion and heatedto reflux for 16 hours. The reaction mixture was cooled to roomtemperature and extracted between ether and cold 1 M NaOH (aqueous). Theorganic fraction was dried over MgSO4 and concentrated in vacuo toafford a dark solid which was subjected to silica gel flash columnchromatography (eluent 100% DCM to 97:3 DCM/diethyl ether), yielding0.91 g of 2-(4-ethyl-2-thiazolyl)-2,3-dihydro-1H-indole (intermediate4).

EXAMPLE A.123-(2-oxo-2-phenyl-ethylcarbamoyl)-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert butyl ester

3,4-Dihydro-1H-isoquinoline-2,3-dicarboxylic acid-2-tertbutyl ester(2.77 g, 10 mmol) and 2-amino-1 phenyl-ethanone (1.71 g, 10 mmol), andHOBT (1-hydroxybenzo-triazole) (2.70 g, 20 mmol) were dissolved indichloromethane (100 ml). The solution was cooled to 0° C. and then(4-dimethylamino-butyl)-ethyl-carbodiimide (2.29 g, 12 mmol) was addedfollowed by NMM (N-methyl-morpholine) (1.31 g, 13 mmol).

The reaction mixture was then warmed to room temperature. After 72 hoursthe reaction mixture was extracted with water, and the organic phaseextracted consecutively with saturated NaHCO₃, 2N citric acid andNaHCO₃, dried over MgSO₄, filtered and concentrated to yield the titleproduct as a yellow foam. Liquid chromatography (LC) indicated thecompound was 86% pure (214 nm), and was used without furtherpurification.

EXAMPLE A.12a

Dehydration of3-(2-oxo-2-phenyl-ethylcarbamoyl)-3,4-dihydro-1H-isoquinoline-2carboxylicacid benzyl ester (prepared in a similar manner as3-(2-oxo-2-phenyl-ethylcarbamoyl)-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert butyl ester of Example A.12) with POCl₃ yields the followingintermediate compound:

The CBZ group is readily removed from the resulting oxazole by treatmentwith iodotrimethylsilane. The resulting nor-amine oxazole intermediatecan be carried on to compound 170 following similar procedures asdescribed for its analogous imidazole intermediates.

EXAMPLE A.133-(4-phenyl-1H-imidazol-2-yl)-3,4,-dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester

The product prepared in Example A.12 above (3.55 g, 9 mmol), NH₄OAc(ammonium acetate) (20.8 g, 270 mmol) and AcOH (acetic acid) (30 mL)were combined at room temperature and the reaction mixture was warmed ona steam bath for about 3 hours. The reaction mixture was then cooled toroom temperature and poured into an ice slurry mix (400 g). To thismixture was added concentrated ammonium hydroxide (50 mL) and ethylether. The layers were separated, and the aqueous phase washed with asecond portion of ethyl ether. The organic phases were combined, driedover MgSO₄, filtered, and concentrated under reduced pressure to yield abrown foam. This sample was purified by preparative HPLC to yield thepurified title compound as a white powder. LC indicated the sample was96% pure at 214 nm. Measured MW (MH⁺): 376

EXAMPLE A.143-(4-phenyl-1H-imidazol-2-yl)-1,2,3,4-tetrahydro-isoquinoline

Triflouroacetic acid (TFA) (4 mL) was cooled in a test tube to about 0°C. To the cool solvent was then added the product prepared in ExampleA.13 (0.75 g, 2 mmol) above. The reaction mixture was allowed to warm toroom temperature over about 45 minutes. Excess TFA was removed under astream of N₂ gas. The residue was partitioned between dichloromethane(15 mL) and saturated NaHCO₃. The aqueous phase was then re-extractedwith a second portion of dichloromethane and the organic phasescombined, dried over MgSO₄ and filtered, to yield the title compound indichloromethane solution. The filtrate was used in the next step(Example A.15) without further purification or isolation. Measured MW(ME⁺): 276

EXAMPLE A.15[1-(4-tert-butoxy-benzyl)-2-oxo-2-[3-(4-phenyl-1H-imidazol-2-yl)-3,4-dihydro-1H-isoquinolin-2-yl]-ethyl]-carbamicacid tert-butyl ester

2-Tert-butoxycarbonylamino-3-(4-tert-butoxy-phenyl)-propionic acid (0.74g, 2.2 mmol) was dissolved in dichloromethane (40 mL) and the reactionmixture cooled to about 0° C. To the solution was then added NMM (0.21g, 2.1 mmol) followed by isobutyl chloroformate (0.27 g, 2 mmol, 0.26mL) and the solution was allowed to stand for about 1.25 hours. To thereaction mixture was then added the product prepared in Example A. 14(0.55 g, 2 mmol) and the reaction mixture stirred for about 16 hours.The reaction mixture was then extracted with water, saturated NaHCO₃, 2Ncitric acid, saturated NaHCO₃, dried over MgSO₄, filtered andconcentrated to yield the title product as a foam. Measured MW (MH⁺):595.

A bromine can be introduced at the 5-position of the imidazole moiety ofthis intermediate compound by reacting said intermediate compound with 1equivalent of Br₂ at 0° C. in chloroform.

A chlorine can be introduced at the 5-position of the imidazole moietyof this intermediate compound by reacting said intermediate compoundwith N-chloro-succinimide.

EXAMPLE A.163-(5-methyl-4-phenyl-1H-imidazol-2-yl)-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester

3-Formyl-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester(1.83 g, 7 mmol) was combined with AcOH (25 mL) to which was immediatelyadded 1-phenyl-propane-1,2-dione (3.11 g, 21 mmol) and NH₄OAc (13.49 g,175 mmol). The reaction mixture was then placed on a steam bath andheated under an argon atmosphere for 20 minutes. The reaction mixturewas cooled in an ice bath and then added to an ice slurry (44 g). Theresulting mixture was basified by addition of concentrated NH₄OH (50 mL)and then extracted twice with diethyl ether (150 mL each). The combinedorganic phases were dried over MgSO4, filtered and concentrated to yieldcrude product. This material was purified by preparative HPLC to yieldthe title compound as a white solid. Measured MW (MH⁺): 390

EXAMPLE A.173-(5-methyl-4-phenyl-1H-imidazol-2-yl)-3,4,-dihydro-1H-isoquinoline

To a solution of TFA (5 mL) cooled to about 0° C. was added the compoundprepared in Example A.16 (1.10 g, 2.82 mmol) and the reaction mixturestirred for about 30 minutes. The reaction mixture was then removed fromthe ice bath and allowed to warm to room temperature. Excess TFA wasremoved under a stream of N₂. The residue was partitioned betweensaturated NaHCO₃ and dichloromethane. The aqueous phase was washed witha second portion of dichloromethane and the organic phases combined. Thecombined organic phase was dried over Na₂SO₄, then filtered to yield thetitle product as a solution in dichloromethane, which was used withoutfurther purification or isolation.

EXAMPLE A.18[1-(4-tert-butoxy-benzyl)-2-[3-(5-methyl-4-phenyl-1H-imidazol-2-yl)-3,4-dihydro-1H-isoquinolin-2-yl]-2-oxo-ethyl]-carbamicacid tert-butyl ester

2-Tert-butoxycarbonylamino-3-(4-tert-butoxy-phenyl)-propionic acid (0.74g, 2.2 mmol) was dissolved in dichloromethane (60 mL), cooled to about0° C. To the reaction mixture was then added NMM (0.30 g, 2.97 mmol),followed by isobutyl chloroformate (0.39 g, 2.82 mmol, 0.37 mL). Thesolution was allowed to stand at 0° C. for about 90 minutes. To thereaction mixture was then added the product prepared in Example A.17(2.82 mmol) as a solution in dichloromethane. The reaction mixture wasthen warmed to room temperature. After 16 h the reaction mixture wasextracted sequentially with water, saturated NaHCO₃, 2N citric acid,saturated NaHCO₃, then dried over MgSO₄, filtered and concentrated toyield crude product. This material was purified via preparative HPLC toyield the title product as a whitish foam. Measured MW (MH⁺): 609

B. Preparation of the Final Compounds

EXAMPLE B.1

4-Methylmorpholine (0.003 mol) was added to intermediate (5) (0.003 mol)dissolved in chloroform (80 ml). After cooling to 0° C., intermediate(25) (0.003 mol) was added neat as an oil. After 27 minutes, thereaction mixture was washed with water, saturated NaHCO₃, and brine,dried, filtered, and concentrated, yielding[2S-[1(R*),2R*]]-1,1dimethylethyl[1-[[2,3-dihydro-2-(4-propyl-1H-imidazol-2-yl)-1H-indol-1-yl]-carbonyl]propyl]-carbamate(compound 14).

EXAMPLE B.2

To intermediate (3) (0.047 mole) in methanol (200 ml) was addedpotassium acetate (0.199 mole). The mixture was heated to reflux underargon. To this was slowly added a solution of 1-amino-2-pentanonehydrochloride (0.094 mole) in methanol (95 ml) over 45 minutes. Afterthe addition was complete, the mixture was allowed to stir overnight atreflux, then concentrated. The concentrate was taken up in DCM andwashed with saturated NaHCO₃. The aqueous layer was extracted with DCM.The combined organic extracts were dried and concentrated to a solidresidue. The residue was purified by trituration with diethyl ether andACN and optionally further purified by column chromatography, yielding5.83 g of(S)-1-acetyl-2,3-dihydro-2-(4-propyl-1H-imidazol-2-yl)-1H-indole(compound 8, mp. 174–175° C.).

EXAMPLE B.3

Intermediate (12) (0.00101 mole), 2–3-hexanedione (0.004 mole), andammonium acetate (0.025 mole) were combined in acetic acid (4 ml), andimmediately placed on a steam bath for 15 minutes. After 2 hours at roomtemperature, the reaction was poured into ice water (100 ml), basifiedwith 3N NaOH, and extracted with diethylether (twice). The organicphases were combined, dried, filtered, and concentrated. The residue wastaken up in diethylether, concentrated and then purified by prep LC,yielding 0.440 g of 1,1-dimethylethyl2,3-dihydro-2-(5-methyl-4-propyl-1H-imidazol-2yl)-1H-indole-1-carboxylate(compound 99).

Analogously, compound (80) was prepared by reacting intermediate (12)with the respective aldehyde of 1,1,1-trifluoro-3,3-dibromoacetone.

EXAMPLE B.4

N-[(1,1-dimethylethoxy)carbonyl]-N-methyl-L-alanine (0.00181 mol) wasdissolved in DCM and cooled to 0° C. Triethylamine (0.00181 mol), thenisobutyl chloroformate (0.00181 mol) were added, and the mixture wasstirred at 0° C. for 70 minutes. Intermediate (5) (0.00181 mol) in DCM(6 ml) was added. The mixture was allowed to warm to room temperatureand stirred overnight. The mixture was extracted (water, saturatedNaHCO₃), dried, filtered, and concentrated. The residue was purified byHPLC. The pure fractions were collected and the solvent was evaporated,yielding 0.380 g of [2S-[1(R*),2R*]]-1,1-dimethylethyl[2-[2,3-dihydro-2-(4-propyl-1H-imidazol-2-yl)-1H-indol-1-yl]-1-methyl-2-oxoethyl]methyl-carbamate(compound 63, mp. 77–80° C.).

EXAMPLE B.5

Compound 14 (0.0073 mole) and trifluoroacetic acid (5 ml), bothprecooled in an ice bath, were combined and allowed to slowly return toroom temperature under nitrogen. After 1 hour, the mixture wasconcentrated. The concentrate was dissolved in water and extracted withdiethylether. The aqueous layer was basified with saturated NaHCO₃ andextracted twice with chloroform. The combined organic extracts weredried over MgSO₄ and concentrated. The residue was dissolved in etherand treated with 3 ml of 1M HCl in ether. The precipitate was filteredand dried under vacuum. The residue was partitioned between saturatedNaHCO₃ and chloroform. The organic layer was dried over MgSO₄ andconcentrated. The concentrate was purified on a Biotage column, elutingwith 5% MeOH in chloroform. The residue was dissolved in ether andtreated with ±2 ml of 1M HCl in diethyl ether. The solid was collectedby filtration under nitrogen and dried under vacuum overnight, yielding0.364 g of[2S-[1(R*),2R*]]-α-ethyl-2,3-dihydro-β-oxo-2-(4-propyl-1H-imidazol-2-yl)-1H-indole-1-ethanaminedihydrochloride dihydrate (compound 15, mp. 132–140° C.).

EXAMPLE B.6

A suspension of intermediate (13) (0.0102 mole) in n-butanol (200 ml)was treated with butanoic acid hydazide (0.0254 mole), stirred for 10minutes, and then heated to reflux for 10 days. The reaction was cooled,concentrated in vacuo, distributed between DCM and distilled water. Theconcentrated organic phase was subjected to reverse phase preparatorycolumn chromatography to give1-acetyl-2,3-dihydro-2-(5-propyl-1H-1,2,4triazol-3-yl)-1H-indole(compound 91).

EXAMPLE B.7

a) A solution of the compound 91 (0.42g) in ethanol (25 ml) was treatedwith an aqueous NaOH solution (3 M, 25 mL) and the reaction mix wasrefluxed for 24 hours. The reaction was cooled, diluted with ethylacetate, and treated with cold distilled water. The layers wereseparated and the aqueous fraction was extracted 5 times with ethylacetate and the combined organic fractions were dried, concentrated andpurified by preparatory column chromatography yielding2,3-dihydro-2-(5-propyl-1H-1,2,4-triazol-3-yl)-1H-indole.

b) A solution of2,3-dihydro-2-(5-propyl-1H-1,2,4-triazol-3-yl)-1H-indole (0.00017 mole)in DCM (5 ml) was treated with N-ethyl-N-(1-methylethyl)-2-propanamine(0.00072 mole) then (2-fluoro-2-oxoethyl)-9H-fluoren-9-yl-carbamic acidmethyl ester (0.00070 mole). The reaction was stirred at roomtemperature for 15 hours. The reaction was diluted with DCM, treatedtwice with saturated NaHCO₃, and dried over Na₂SO₄ and concentrated. Theresidue was subjected to reverse phase prep column chromatography toobtain 0.02 g of the desired mono-adduct and 0.02 g of a bis-adduct thatwas completely converted to the desired mono-adduct by treatment withthe prep chromatography eluent (0.1% trifluoroacetic acid inwater/acetonitrile). These were combined, yielding 0.03 g ofH-fluoren-9-ylmethyl[2-[2,3-dihydro-2-(5-propyl-1H-1,2,4-triazol-3-yl)-1H-indol-1-yl]-2-oxoethyl]-carbamate.

c) A solution of H-fluoren-9-ylmethyl[2-[2,3-dihydro-2-(5-propyl-1H-1,2,4-triazol-3yl)-1H-indol-1-yl]-2-oxoethyl]-carbamate(0.00006 mole) in DCM (10 ml) was treated with piperidine (0.010 mole)and stirred at room temperature for 1 hour. The completed reaction wasconcentrated in vacuo and subjected to reverse phase prep columnchromatography, yielding 0.02 g of2,3-dihydro-β-oxo-2-(5-propyl-1H-1,2,4-triazol-3-yl)-1H-indole-1-ethanaminetrifluoroacetate (1:1) (compound 92).

EXAMPLE B.8

A mixture of intermediate (14) (0.00898 mole) and butanoyl chloride(0.0094 mole) in pyridine (140 ml) was stirred at room temperature for40 hours and then heated to reflux. After 21 hours the reaction wascooled and concentrated in vacuo. The residue was extracted between DCMand saturated aqueous NaHCO₃ and the organic fraction was dried overNa₂SO₄, filtered, and concentrated.The residue was subjected to silicagel flash column chromatography (eluent 100% CH₂Cl₂ to 95/5CH₂Cl₂/ether), yielding1-acetyl-2,3-dihydro-2-(5-propyl-1,2,4-oxadiazol-3-yl)-1H-indole(compound 89, mp. 93–94° C.).

EXAMPLE B.9

a) A solution of compound 89 (0.0035 mole) in ethanol (60 ml) wastreated with 3M NaOH (60 ml), and the reaction mix was heated to 55–60°C. for 5.5 hours. The reaction was rapidly cooled in an ice bath,diluted with DCM, and treated with cold distilled water. The layers wereseparated and the aqueous fraction was extracted three times with DCM.The organic fractions were combined, washed once with 1M NaOH, and driedover Na₂SO₄ and concentrated in vacuo. The residue was purified by prepcolumn chromatography, yielding 0.45 g of2,3-dihydro-2-(5-propyl-1,2,4-oxadiazol-3yl)-1H-indole.

b) A solution of 2,3-dihydro-2-(5-propyl-1,2,4-oxadiazol-3-yl)-1H-indole(0.0011 mole) in DCM (10 ml)was treated withN-methyl-N-(1-methylethyl)-2-propanamine (0.40 mL) then(2-fluoro-2-oxoethyl)-9H-fluoren-9-yl carbamic acid methyl ester (0.67g). The reaction was stirred at room temperature for 40 hours andtreated with another portion each ofN-methyl-N-(1-methylethyl)-2-propanamine then(2-fluoro-2-oxoethyl)-9H-fluoren-9-yl carbamic acid methyl ester andstirred at room temperature for two days. The reaction was diluted withDCM, treated twice with saturated NaHCO₃, and dried over Na₂SO₄. andconcentrated. The residue was subjected to reverse phase prep columnchromatography, yielding 0.35 g of 9H-fluoren-9-ylmethyl[2-[2,3-dihydro-2-(5-propyl-1,2,4-oxadiazol-3-yl)-1H-indol-1-yl]-2-oxoethyl]-carbamate.

c) 9H-fluoren-9-ylmethyl [2-[2,3-dihydro-2-(5-propyl-1,2,4-oxadiazol-3-yl)-1H-indol-1-yl]-2-oxoethyl]-carbamate (0.35 g) wasdissolved in DCM (40 ml), treated with piperidine (0.50 ml), and stirredat room temperature for 18 hours. The completed reaction wasconcentrated in vacuo and subjected to reverse phase prep columnchromatograph, yielding 0.13 g of2,3-dihydro-β-oxo-2-(5-propyl-1,2,4-oxadiazol-3-yl)-1H-indole-1-ethanaminetrifluoroacetate (1:1) (compound 90, mp. 160–162° C.).

EXAMPLE B.10

2,3-dihydro-2-(4-propyl-1H-imidazol-2-yl)-1H-indole (0.0024 mol) and1,3-isobenzofurandione (0.0026 mol) were heated to 100° C. in a 25 mlpear shaped flask under argon for 2 hours. The mixture was dissolved inmethanol and heated to reflux for 15 hours. The reaction mixture wasconcentrated and taken up in DCM, washed with water and 3 N NaOH. Thebasic aqueous extract was acidified with 6 N HCl and extracted with DCM.This organic extract was dried over MgSO₄ and concentrated. Theconcentrate was triturated in ether and collected. This was furtherpurified, together with the acidic aqueous solution, by prep liquidchromatography, yielding 0.23 g of2-[[2-(4-ethyl-1H-imidazol-2-yl)-2,3-dihydro-1H-indol-1-yl]carbonyl]-benzoicacid trifluoroacetate (1:1) (compound 85, mp. 98–103° C.).

EXAMPLE B.11

1-Isocyanato-2-nitro-benzene (0.002 mol) was added to a solution ofintermediate (15) (0.016 mol) in THF (10 ml). The mixture was stirred atroom temperature under argon for 5 hours. The mixture was diluted withhexanes, filtered, and allowed to dry, yielding 0.34 g of2-(4-ethyl-1H-imidazol-2-yl)-2,3-dihydro-N-(2-nitrophenyl)-1H-indole-1-carboxamide(compound 77, mp. 208–209° C.).

EXAMPLE B.12

To a mixture of compound 77 (0.0006 mol), Raney Nickel (0.02 g; 50%slurry in water), and methanol (20 ml) was added hydrazin. Water (0.003mol). The resulting mixture was heated to reflux for 2 hours. Aftercooling to room temperature, the mixture was carefully filtered throughcelite and the filtrate was concentrated. The residue was triturated inether and filtered. The residue was purified by prep liquidchromatography, yielding 0.24 g ofN-(2-aminophenyl)-2-(4-ethyl-1H-imidazol-2-yl)-2,3-dihydro-1H-indole-1-carboxamidetrifluoroacetate (1:2) (compound 79, mp. 106–108° C.).

EXAMPLE B.13

A mixture of compound 16 (0.00697 mole) in THF (70 ml) was treated withof sodium hydride (0.007 mole) in one portion and stirred at ambienttemperature for 16 hours. Iodomethane (0.0071 mole) was introduced inone portion. After stirring at ambient temperature for 24 hours, moresodium hydride (0.007 mole) was added in one portion under an argonatmosphere. The flask was restoppered after effervescence had subsided,and stirred for 16 hours. The completed reaction was cooled in an icebath, poured into DCM, and treated with cold water. The layers wereseparated and the aqueous was extracted three times with DCM. Thecombined organic fractions were washed with sat NaHCO₃, dried overNa₂SO₄, and concentrated. The residue was subjected to flash silica gelcolumn chromatography (DCM to ether to 9:1 ether/THF). The appropriatefractions were combined. The residue was taken up in ether and placed inthe freezer. Crystallization occurred, yielding 0.55 g (29.3%) of1-acetyl-2-(4-ethyl-1-methyl-1H-imidazol-2-yl)-2,3-dihydro-1H-indole(compound 132, mp. 105–106° C.). The second set of fractions werecombined. The residue was taken up in ether and placed in the freezer.Observed crystallization occurred, yielding 0.38 g of1-acetyl-2-(4-ethyl-1-methyl-1H-imidazol-2-yl)-2,3-dihydro-1H-indole(compound 133, mp. 135–137° C.).

EXAMPLE B.14

Compound 80 (0.001 mole) was suspended in 1N NaOH (12 ml). The mixturewas vigorously stirred and heated to 88° C. under nitrogen for 1 hour.After stirring at room temperature for 3 hours, the mixture was cooledto 0° C., slowly neutralized with 1M HCl to precipitate some solid. Thesolid was filtered, rinsing with ice cold water. The aqueous phase wasextracted twice, dried, filtered, concentrated and dried, yielding 0.140g of 1,1-dimethylethyl2-(4-carboxy-1H-imidazol-2-yl)-2,3-dihydro-1H-indole-1-carboxylate(compound 117).

EXAMPLE B.15

1-Hydroxybenzotriazole hydrate (0.00036 mole), glycine methylester,hydrochloride (0.00047 mole), 4-methylmorpholine (0.00055 mole), andN′-(ethylcarbonimidoyl)-N,N-dimethyl-1,3-propanediaminemonohydrochloride (0.00047 mole) were added to compound 117 (0.00036mole) dissolved in DCM (30 ml) at 0° C. The mixture was allowed to warmto room temperature under nitrogen, and stirred overnight. The mixturewas extracted with water, saturated NaHCO₃, 2N citric acid, thensaturated NaHCO₃, dried, filtered, and concentrated, yielding 0.100 g(69%) of 1,1-dimethylethyl2,3-dihydro-2-[4-[[(2-methoxy-2-oxoethyl)amino]carbonyl]-1H-imidazol-2-yl]-1H-indole-1-carboxylate(compound 118).

EXAMPLE B.16

Compound 61 (0.00028 mol) was treated with 3N NaOH (3 ml) and allowed tostir for 20 minutes at room temperature. The solution was then treatedwith 3 ml of 3 N HCl and extracted with chloroform. The material stayedin the aqueous layer. The aqueous layer was purified by preparativeliquid chromatography, yielding 0.12 g of2-[1-(aminoacetyl)-2,3-dihydro-1H-indol-2-yl]-1H-benzimidazole-5-carboxylicacid monohydrate trifluoroacetate (1:2) (compound 62, mp. 208–211° C.).

EXAMPLE B.17

Compound 102 (0.00238 mole) was dissolved in 40 ml of methanol andcombined with 1N KOH (50 mL). The reaction was warmed to 40° C. underargon overnight. The heat was increased to 55–60° C. for an additionalovernight heating. The reaction was then cooled to room temperature,filtered, and at 0° C. slowly neutralized with 1N HCl. The sample wasextracted 5 times with DCM, combined, and dried over Na₂SO₄. Thisorganic solution was filtered and used in further synthesis withoutfurther purification, yielding 1,1-dimethylethyl2-(4-carboxy-5-propyl-1H-imidazol-2-yl)-2,3-dihydro-1H-indole-1-carboxylate(compound 105).

EXAMPLE B.18

1-Hydroxybenzotriazole hydrate (0.00318 mole) was added to a solution ofcompound 105 (0.00159 mole) in DCM (160 mL) at room temperture.N,N′-methane-tetrayl-biscyclohexanamine (0.00206 mole) was added neat atroom temperature. After 60 minutes, NH₃ gas was bubbled in for 5minutes, and a solid precipitated out. The mixture was allowed to sitover the weekend. The mixture was filtered, and the filtrate wasextracted with saturated NaHCO₃. The organic phases were dried overMgSO₄, filtered, and concentrated. The residue was purified by liquidchromatography, yielding 0.21 g of 1,1-dimethylethyl2-[4-(aminocarbonyl)-5-propyl-1H-imidazol-2-yl]-2,3-dihydro-1H-indole-1-carboxylate(compound 106).

EXAMPLE B.19

1-Hydroxybenzotriazole hydrate (0.00158 mole) was added to a solution ofcompound 105 (0.00079 mole) in DCM (80 ml). Glycine methylesterhydrochloride (0.00103 mole), N′-(ethylcarbonimidoyl)-N,N-dimethyl-1,3propanediamine monohydrochloride (0.00103 mole) and 4-methylmorpholine(0.00103 mole) were added. THF (25 mm) was added. The reaction wasstirred at room temperature for 3 days. The mixture was extracted withwater. The organic phase was washed with saturated NaHCO₃, 2N citricacid, saturated NaHCO₃, dried over MgSO₄, filtered, and concentrated,yielding 0.20 g of 1,1-dimethylethyl2,3-dihydro-2-[4-[[(2-methoxy-2-oxoethyl)amino]carbonyl]-5-propyl-1H-imidazol-2-yl]-1H-indole-1-carboxylate(compound 109).

EXAMPLE B.20

Compound 81 (0.0005 mole) was suspended in 1N NaOH (6 ml) under argon.The mixture was immediately heated to 80° C. for 60 minutes. At roomtemperature, chloroform (6 ml) then(2-fluoro-2-oxoethyl)-1,1-dimethylethyl carbamic acid ester (0.001 mole)were added. The mixture was stirred overnight. The layers wereseparated. The aqueous phase was cooled, acidified, and extracted twicewith chloroform. The latter organic phases were combined, dried,filtered, and concentrated. The sample was purified by prep HPLC,yielding 0.040 g of2-[1-[[[(1,1-dimethylethoxy)carbonyl]-amino]acetyl]-2,3-dihydro-1H-indol-2-yl]-1H-imidazole4-carboxylicacid (compound 138).

EXAMPLE B.21

To compound 145 (0.00097 mole), dissolved in ethanol (5 ml), was addedseveral drops of 21% NaOEt in ethanol. The mixture was allowed to stirat room temperature under argon. An additional 2 drops of 21% NaOEt inethanol were added after 30 minutes. An additional 2 drops of 21% NaOEtin ethanol were added after 16 hours. After 30 minutes the mixture wasconcentrated and partitioned between water and DCM. The aqueous layerwas washed with additional DCM. The combined organics were washed withwater, dried, and concentrated, yielding 0.193 g (66%) of[2S-[1(R*),2R*]]-2,3-dihydro-α-methyl-β-oxo-2-(4-propyl-1H-imidazol-2-yl)-1H-indole-1-ethanol(compound 146).

Compound 148 was prepared analogously starting from compound 147.

EXAMPLE B.22

To a suspension of compound 58 (0.0019 mole) in acetonitrile (15 ml) wasadded acetic acid, anhydride (0.074 mole). Stirred at room temperatureunder argon for 4 hours. An additional 1.0 ml of acetic acid, anhydridewas added, and the reaction was stirred overnight. After stirring 6hours more, the reaction was complete. The mixture was concentrated andthe residue partitioned between saturated NaHCO₃ and chloroform. Theorganic layer was dried and concentrated. The residue was purified bycolumn chromatography. The desired fractions were combined, trituratedin ether and collected. yielding 0.37 g of1-[[1-[(4-chlorophenyl)acetyl]4-(3-methoxyphenyl)4-piperidinyl]methyl]-1,3-dihydro-2H-benzimidazol-2-one(compound 149).

EXAMPLE B.23

A solution of compound 149 (0.0012 mole) and THF (200 ml) was placedinside of a photochemical reactor and irradiated with UV light for 14hours. The mixture was then allowed to sit at room temperature undernitrogen for 2 days. The mixture was concentrated. The concentrate waspurified on Biotage column, eluting with 1:9 THF in DCM, yielding 0.077g of1-[2-(1-acetyl-2,3-dihydro-1H-indol-2-yl)-5-propyl-1H-imidazol-4-yl]-ethanone(compound 150).

EXAMPLE B.24

Compound 13 (0.00106 mole) dissolved in 10 ml of THF was treated at roomtemperature with BH₃.THF (19 ml), which was a solution in THF. Thesolution was then placed in an oil bath and heated to 60° C. overnight.After cooling to 0° C., the solution was carefully treated with 15 ml of3N HCl. The reaction was then warmed to room temperature and stirred for4 hours. The mixture was then recooled to 0° C. and basified with 12 mlof 3N NaOH, then completion of basification was done with solid Na₂CO₃.The layers were separated and the aqueous was rewashed with chloroform.The organics were combined, a small amount of aqueous separated, and theorganic dried over Na₂SO₄. The mixture was filtered, and the filtrateconcentrated under reduced pressure. The residue was submitted forpreparative liquid chromatography, yielding 0.33 g of[2S-[1(R*),2R*]]-2-(4-ethyl-1H-imidazol-2-yl)-2,3-dihydro-α-methyl-1H-indole-1-ethanaminetrifluoroacetate (1:1) (compound 127).

EXAMPLE B.253-amino-4-(4-hydroxy-phenyl)-1-[3-(4-phenyl-1H-imidazol-2-yl)-3,4-dihydro-1-H-isoquinolin-2-yl]-butan-1-one(compound 155)

TFA (4 mL) was cooled to about 0° C. and then the product prepared inExample A.15 (1.10 g, 1.85 mmol) was added. The reaction mixture sat forabout 0.5 hours. Excess TFA was then removed under a stream of N₂ toyield a brown oil. The oil was purified via preparative HPLC to yieldthe title compound as a white solid. Measured MW (MH⁺): 439

EXAMPLE B.262-amino-3-(4-hydroxy-benzyl)-1-[3-(5-methyl-4-phenyl-imidazol-2-yl)-3,4-dihydro-1H-isoquinolin-2-yl]-propan-1-one(compound 153)

To a solution of TFA (4mL) cooled to about 0° C. was added the compoundprepared in Example A. 18 (0.24 g, 0.4 mmol) and the reaction mixturestirred for about 20 minutes. The reaction mixture was then removed fromthe ice bath and allowed to warm to room temperature. Excess TFA wasremoved under a stream of N₂ to yield crude product. This material waspurified via preparative HPLC to yield the title compound as a whitesolid.

Measured MW (MH⁺): 453

Table F-1 lists the compounds that were prepared according to one of theabove Examples. The following abbreviations were used in the tables:.C₂HF₃O₂ stands for the trifluoroacetate salt, .2C₂H₂O₄ stands for theethanedioate salt, and .C₁₀H₈O₃S stands for the 2-naphthalenesulfonatesalt. Said Table F-1 lists the structure of the compounds, the Examplenumber according to which these compounds have been prepared, the saltform, the stereochemical designation and the melting point (ifmeasured).

TABLE F-1

Co. No. 1; Ex. B.1

Co. No. 2; Ex. B.5

Co. No. 3; Ex. B.1; [2R-[1(S*), 2R*]] + [2S-(1(R*), 2R*]]

Co. No. 4; Ex. B.1; [1(S), 2A]

Co. No. 5; Ex. B.1; [1(S), 2B]

Co. No. 6, Ex. B.5; [1(S), 2A]

Co. No. 7; Ex. B.5; [1(S), 2B]

Co. No. 8; Ex. B.2; (S); mp. 174–175° C.

Co. No. 9; Ex. B.1; [2S-[1(R*), 2R*]]

Co. No. 10; Ex. B.5; [2S-[1(R*), 2R*]]

Co. No. 11; Ex. B.2; (S); mp. 136–139° C.

Co. No. 12; Ex. B.1; [2S-[1(R*), 2R*]]

Co. No. 13; Ex. B.5; [2S-[1(R*), 2R*]]; mp. 116–118° C.

Co. No. 14; Ex. B.1; [2S-[1(R*), 2R*]]

Co. No. 15; Ex. B.5; .2HCl.2H₂O [2S-[1(R*), 2R*]] mp. 132–140° C.

Co. No. 16; Ex. B.2

Co. No. 17; Ex. B.1; [2R-[1(S*), 2R*]]; mp. 76–79° C.

Co. No. 18; Ex. B.1; mp. 198–199° C.

Co. No. 19; Ex. B.5; mp. 184–186° C.

Co. No. 20; Ex. B.5; .H₂O [2R-[1(S*), 2R*]]; mp. 73–74° C.

Co. No. 21; Ex. B.1; [2R-[1(S*), 2R*]] + [2S-[1(R*), 2R*]]

Co. No. 22; Ex. B.1 [1(S), 2A]

Co. No. 23; Ex. B.1; [1(S), 2B]

Co. No. 24; Ex. B.5; .2C₂H₂O₄; [1(S), 2A]; mp. >90° C.

Co. No. 25; Ex. B.5; .2HCl.2H₂O [1(S), 2B]; mp. >100° C.

Co. No. 26; Ex. B.2; (S); mp. 208–210° C.

Co. No. 27; Ex. B.4; [S-[1(R*), R*]; mp. 107–109° C.

Co. No. 28; Ex. B.5; .3HCl; [S-[1(R*), R*]; mp. 240–242° C.

Co. No. 29; Ex. B.4; mp. 170–171° C. Co. No. 30; Ex. B.4; .C₂HF₃O₂;[S-(R*, R*)]; mp. 173–175° C.

Co. No. 31; Ex. B.2; mp. 261–262° C.

Co. No. 32; Ex. B.2; mp. 256–257° C.

Co. No. 33; Ex. B.1; .C₂HF₃O₂; [R-(R*, S*)]

Co. No. 34; Ex. B.1; .C₂HF₃O₂; [S-(R*, R*)]

Co. No. 35; Ex. B.1; .C₂HF₃O₂; [R-(R*, S*)] + [S-(R*, R*]]

Co. No. 36; Ex. B.5; .2C₂HF₃O₂; [S-(R*R*)]

Co. No. 37; Ex. B.5; .2C₂HF₃O₂; [R-(R*, S*)] + [S-(R*, R*)]

Co. No. 38; Ex. B.2; mp. 214–216° C.

Co. No. 39; Ex. B.1; [1(S)]; mp. 137–138° C.

Co. No. 40; Ex. B.5; [1(S)]; mp. 198–203° C.

Co. No. 41; Ex. B.2; mp. 221–222° C.

Co. No. 42; Ex. B.1; .2C₂HF₃O₂; [R-(R*, S*)]

Co. No. 43; Ex. B.1 .C₂HF₃O₂; [S-(R*, R*)]

Co. No. 44; Ex. B.5; .3C₂HF₃O₂; [S-(R*, R*)]

Co. No. 45; Ex. B.5; .2C₂HF₃O₂; [R-(R*, S*)] + [S-(R*, R*)]

Co. No. 46; Ex. B.5; mp. 158–160° C.

Co. No. 47; Ex. B.5; .2C₂H₂O₄; [S-(R*, R*)] mp. 135–137° C.

Co. No. 48; Ex. B.5; .2HCl.3H₂O; [S-(R*, R*)] mp. 85–87° C.

Co. No. 49; Ex. B.4; .H₂O.C₂HF₃O₂; mp.

Co. No. 50; Ex. B.5; mp. 116–118° C.

Co. No. 51; Ex. B.1; .C₂HF₃O₂; [R-(R*, S*)] + [S-(R*, R*)]

Co. No. 52; Ex. B.5; .2C₂HF₃O₂; [R-(R*, S*)]

Co. No. 53; Ex. B.5; .H₂O.2C₂HF₃O₂; [R-(R*, S*)]

Co. No. 54; Ex. B.4; .C₂HF₃O₂; [S-(R*, R*)] mp. 66–68° C.

Co. No. 55; Ex. B.5; .C₁₀H₈O₃S.H₂O; [S-(R*, R*)]; mp. 195–197° C.

Co. No. 56; Ex. B.1; [S-(R*, R*)]; mp. 76–78° C.

Co. No. 57; Ex. B.5; [S-(R*, R*)]; mp. 141–143° C.

Co. No. 58; Ex. B.2; mp. 173–174° C.

Co. No. 59; Ex. B.2; mp. 220–222° C.

Co. No. 60; Ex. B.1; .H₂O; mp. 183° C.

Co. No. 61; Ex. B.5; mp. 122° C.

Co. No. 62; Ex. B.16; .2H₂O.2C₂HF₃O₂

Co. No. 63; Ex. B.4; [S-(R*, R*)]; mp. 77–80° C.

Co. No. 64; Ex. B.5; [S-(R*, R*)]; mp. 137–138° C.

Co. No. 65; Ex. B.1; [2S-[1(R*), 2R*]]

Co. No. 66; Ex. B.5; .C₂H₂O₄.2H₂O; [2S-[1(R*), 2R*]]; mp. 153–156° C.

Co. No. 67; Ex. B.1; mp. 100–104° C.

Co. No. 68; Ex. B.5; .HCl.H₂O; mp. 152° C.

Co. No. 69; Ex. B.1

Co. No. 70; Ex. B.5; .H₂O; mp. 168–170° C.

Co. No. 71; Ex. B.2; 2KCl; mp. 189–191° C.

Co. No. 72; Ex. B.1; mp. 168° C.

Co. No. 73; Ex. B.5; .H₂O.2C₂F₃O₂; mp. >300° C.

Co. No. 74; Ex. B.2; mp. 191–192° C.

Co. No. 75; Ex. B.1; mp. 214–216° C.

Co. No. 76; Ex. B.5; mp. 158–160° C.

Co. No. 77; Ex. B.11

Co. No. 78; Ex. B.11

Co. No. 79; Ex. B.12; .2C₂HF₃O₂

Co. No. 80; Ex. B.3; mp. 179–181° C.

Co. No. 81; Ex. B.1

Co. No. 82; Ex. B.5; mp. 186–188° C.

Co. No. 83; Ex. B.1

Co. No. 84; Ex. B.5; .C₄H₄O₄; mp. 173–174° C.

Co. No. 85; Ex. B.10; .C₂HF₃O₂

Co. No. 86; Ex. B.2; mp. 225–226° C.

Co. No. 87; Ex. B.4;

Co. No. 88; Ex. B.5; mp. 193–195° C.

Co. No. 89; Ex. B.8

Co. No. 90; Ex. B.9; .C₂HF₃O₂

Co. No. 91; Ex. B.6; mp.

Co. No. 92; Ex. B.7; .C₂HF₃O₂

Co. No. 93; Ex. B.2

Co. No. 94; Ex. B.1; .C₂HF₃O₂

Co. No. 95; Ex. B.5; .C₂HF₃O₂

Co. No. 96; Ex. B.2

Co. No. 97; Ex. B.4

Co. No. 98; Ex. B.5; mp. 214–215° C.

Co. No. 99; Ex. B.3

Co. No. 100; Ex. B.1; mp. 165–167° C.

Co. No. 101; Ex. B.5; mp. 197–198° C.

Co. No. 102; Ex. B.3

Co. No. 103; Ex. B.4

Co. No. 104; Ex. B.5; .C₂HF₃O₂; mp. 102–105° C.

Co. No. 105; Ex. B.17

Co. No. 106; Ex. B.18

Co. No. 107; Ex. B.1

Co. No. 108; Ex. B.5; .C₂HF₃O₂; mp. 124–131° C.

Co. No. 109; Ex. B.19

Co. No. 110; Ex. B.1

Co. No. 111; Ex. B.5; .C₂HF₃O₂; mp. 95–99° C.

Co. No. 112; Ex. B.2; mp. 236–237° C.

Co. No. 113; Ex. B.1; mp. 184–188° C.

Co. No. 114; Ex. B.5; .C₂HF₃O₂

Co. No. 115; Ex. B.1; .C₂HF₃O₂ [2R-[1(S*), 2R*]]

Co. No. 116; Ex. B.5; .C₂HF₃O₂ [2R-[1(S*), 2R*]]

Co. No. 117; Ex. B.14

Co. No. 118; Ex. B.15

Co. No. 119; Ex. B.1

Co. No. 120; Ex. B.5; .C₂HF₃O₂

Co. No. 121; Ex. B.1; .C₂HF₃O₂ [2S-[1(R*), 2R*]]

Co. No. 122; Ex. B.5; .C₂HF₃O₂ [2S-[1(R*), 2R*]]

Co. No. 123; Ex. B.1; .C₂HF₃O₂ [2R-[1(S*), 2R*]]

Co. No. 124; Ex. B.5; .C₂HF₃O₂ [2R-[1(S*), 2R*]]

Co. No. 125; Ex. B.4; .C₂HF₃O₂ [2R-[1(S*), 2R*]]

Co. No. 126; Ex. B.5; .C₂HF₃O₂ [2R-[1(S*), 2R*]]

Co. No. 127; Ex. B.24; .C₂HF₃O₂; [2S-[1(R*), 2R*]]

Co. No. 128; Ex. B.1; .C₂HF₃O₂; [2S-[1(R*), 2R*]]

Co. No. 129; Ex. B.5; .C₂HF₃O₂; [2S-[1(R*), 2R*]]

Co. No. 130; Ex. B.4; .C₂HF₃O₂; [2S-[1(R*), 2R*]]

Co. No. 131; Ex. B.5; .HCl [2S-[1(R*), 2R*]]; mp. 235–240° C.

Co. No. 132; Ex. B.13

Co. No. 133; Ex. B.13

Co. No. 134; Ex. B.1

Co. No. 135; Ex. B.5; mp. 115–117° C.

Co. No. 136; Ex. B.1

Co. No. 137; Ex. B.5; mp. 107–109° C.

Co. No. 138; Ex. B.20

Co. No. 139; Ex. B.5; .C₂HF₃O₂

Co. No. 140; Ex. B.2

Co. No. 141; Ex. B.1; .C₂HF₃O₂; [2R-[1(S*), 2R*]]

Co. No. 142; Ex. B.5; [2R-[1(S*), 2R*]]

Co. No. 143; Ex. B.1; .C₂HF₃O₂; [2S-[1(R*), 2R*]]

Co. No. 144; Ex. B.5; [2S-[1(R*), 2R*]]

Co. No. 145; Ex. B.1; [2S-[1(R*), 2R*]]

Co. No. 146; Ex. B.21; [2S-[1(R*), 2R*]]

Co. No. 147; Ex. B.1; [2R-[1(S*), 2R*]]

Co. No. 148; Ex. B.21; [2R-[1(S*), 2R*]]

Co. No. 149; Ex. B.22

Co. No. 150; Ex. B.23

Co. No. 151; Ex. B.1

Co. No. 152; Ex. B.5; .C₂HF₃O₂

Co. No. 153; Ex. B.26

Co. No. 154; Ex. B.26

Co. No. 155; Ex. B.25

Co. No. 156; Ex. B.26

Co. No. 157; Ex. B.26

Co. No. 158; Ex. B.26

Co. No. 159; Ex. B.26

Co. No. 160; Ex. B.26

Co. No. 161; Ex. B.25

Co. No. 162; Ex. B.26

Co. No. 163; Ex. B.25

Co. No. 164; Ex. B.25

Co. No. 165; Ex. B.25

Co. No. 166; Ex. B.25

Co. No. 167; Ex. B.25

Co. No. 168; Ex. B.25

Co. No. 169; Ex. B.25

Co. No. 170; Ex. B.25

Co. No. 171; Ex. B.25

Co. No. 172; Ex. B.25

Co. No. 173; Ex. B.26

Co. No. 174; Ex. B.26

Co. No. 175; Ex. B.26

Co. No. 176; Ex. B.26

Co. No. 177; Ex. B.26

Co. No. 178; Ex. B.26

Co. No. 179; Ex. B.26

Co. No. 180; Ex. B.26

Co. No. 181; Ex. B.26C. Pharmacological ExamplesC.1. Inhibition of Tripeptidyl Peptidase II (TPP II)

The inhibition of TPP II was measured using the procedure as describedby C. Rose et al. in Nature, 380, 403–409 (1996).

TPPII activity was evaluated using 15 μM AAF-AMC as a substrate in a 50mM Potassiumphosphate buffer pH 7.5 with 1 mM DTT and 1 mM EGTA.Compounds were added at a final DMSO concentration of 1%. Fluorescencewas measured at 405 nm. The potency of the compounds of formula (I) wasexpressed as the IC₅₀ value, i.e. the concentration needed to provide50% inhibition. Compounds 6, 10, 13, 15, 19, 22, 24, 28, 30, 44, 47, 48,54, 55, 57, 61, 62, 66, 68, 70, 73,76, 82, 84, 88,90,92,95, 101, 104,108, 111, 114, 116, 120, 122, 124, 126, 129, 131, 135, 142, and 144 havean IC₅₀ value equal to or lower than 1.10⁻⁵ M.

C.2 Rat Brain δ-Opioid Receptor Binding Assay

Male, Wistar rats (150–250 g, VAF, Charles River, Kingston, N.Y.) arekilled by cervical dislocation, and their brains removed and placedimmediately in ice cold Tris HCl buffer (50 mM, pH 7.4). The forebrainsare separated from the remainder of the brain by a coronal transection,beginning dorsally at the colliculi and passing ventrally through themidbrain-pontine junction. After dissection, the forebrains arehomogenized in Tris buffer in a Teflon®-glass homogenizer. Thehomogenate is diluted to a concentration of 1 g of forebrain tissue per100 mL Tris buffer and centrifuged at 39,000×G for 10 min. The pellet isre-suspended in the same volume of Tris buffer with several brief pulsesfrom a Polytron homogenizer. This particulate preparation is used forthe δ-opioid binding assays. Following incubation with the δ-selectivepeptide ligand [³H]DPDPE at 25° C., the tube contents are filteredthrough Whatman GF/B filter sheets on a Brandel cell harvester. Thetubes and filters are rinsed three times with 4 mL of 10 mM HEPES (pH7.4), and the radioactivity associated with the filter circles isdetermined using Formula 989 scintillation fluid (New England Nuclear,Boston, Mass.) in a scintillation counter.

The data are used to calculate either the % inhibition compared tocontrol binding (when only a single concentration of test compound isevaluated) or a K_(i) value (when a range of concentrations is tested).

% Inhibition is calculated as follows:

$( {1 - \lbrack \frac{( {{{Test}\mspace{14mu}{Compound}\mspace{14mu}{dpm}} - {{Non}\text{-}{specific}\mspace{14mu}{dpm}}} )}{( {{{Total}\mspace{14mu}{dpm}} - {{Non}\text{-}{specific}\mspace{14mu}{dpm}}} )} \rbrack} ) \times 100\;\%$K_(i) value is calculated using the LIGAND (Munson, P. J. and Rodbard,D., Anal. Biochem. 107: 220–239, 1980) data analysis program.C.3 Rat Brain μ-Opioid Receptor Binding Assay

Male, Wistar rats (150–250 g, VAF, Charles River, Kingston, N.Y.) arekilled by cervical dislocation and their brains removed and placedimmediately in ice cold Tris HCl buffer (50 mM, pH 7.4). The forebrainsare separated from the remainder of the brain by a coronal transection,beginning dorsally at the colliculi and passing ventrally through themidbrain-pontine junction. After dissection, the forebrains arehomogenized in Tris buffer in a Teflon®-glass homogenizer. Thehomogenate is diluted to a concentration of 1 g of forebrain tissue per100 mL Tris buffer and centrifuged at 39,000×G for 10 min. The pellet isre-suspended in the same volume of Tris buffer with several brief pulsesfrom a Polytron homogenizer. This particulate preparation is used forthe μ-opioid binding assays. Following incubation with the μ-selectivepeptide ligand [³H]DAMGO at 25° C., the tube contents are filteredthrough Whatman GF/B filter sheets on a Brandel cell harvester. Thetubes and filters are rinsed three times with 4 mL of 10 mM HEPES (pH7.4) and the radioactivity associated with the filter circles isdetermined using Formula 989 scintillation fluid (New England Nuclear,Boston, Mass.) in a scintillation counter.

The data are used to calculate either the % inhibition compared tocontrol binding (when only a single concentration of test compound isevaluated) or a K_(i) value (when a range of concentrations is tested).

% Inhibition is calculated as follows:

$( {1 - \lbrack \frac{( {{{Test}\mspace{14mu}{Compound}\mspace{14mu}{dpm}} - {{Non}\text{-}{specific}\mspace{14mu}{dpm}}} )}{( {{{Total}\mspace{14mu}{dpm}} - {{Non}\text{-}{specific}\mspace{14mu}{dpm}}} )} \rbrack} ) \times 100\;\%$K_(i) value was calculated using the LIGAND (Munson, P. J. and Rodbard,D., Anal. Biochem. 107: 220–239, 1980) data analysis program.

1. A compound of formula (I)

a stereochemically isomeric form thereof, or a pharmaceuticallyacceptable addition salt thereof, wherein n is [an integer 0] 1; Xrepresents [O; or ]—(CR⁴R⁵)_(m)— wherein m is [an integer] 1or 2; R⁴ andR⁵ are each independently from each other hydrogen or C₁₋₄alkyl; R¹ isC₁₋₆alkylcarbonyl optionally substituted with hydroxy;[C₁₋₆alkylcarbonyl]; aminoC₁₋₆alkylcarbonyl wherein the C₁₋₆alkyl groupis optionally substituted with C₃₋₆cycloalkyl; mono- anddi(C₁₋₄alkyl)aminoC₁₋₆alkylcarbonyl; aminocarbonyl substituted witharyl; C₁₋₆alkylcarbonyloxyC₁₋₆alkylcarbonyl;C₁₋₆alkyloxycarbonylaminoC₁₋₆alkylcarbonyl wherein the amino group isoptionally substituted with C₁₋₄alkyl; an amino acid [residue ] boundvia the carbonyl group; C₁₋₆alkyl substituted with amino; orarylcarbonyl, wherein aryl is phenyl, or phenyl substituted with amino,nitro or hydroxycarbonyl; R² is [a 5-membered heterocycle selected from]

 wherein m′ is an integer 1 to 2; R⁶ is hydrogen or C₁₋₄alkyl; R⁷ isindependently from each other; halo; [amino]; [hydroxy;]trifluoromethyl; C₁₋₆alkyl; phenyl; aminocarbonyl; hydroxycarbonyl; orC₁₋₄alkyloxycarbonyl; [C₁₋₄alkylcarbonyl; orC₁₋₄alkyloxycarbonylC₁₋₄alkylcarbonyl;] or R² is benzimidazole, orbenzimidazole substituted with one or two substituents eachindependently selected from halo, trifluoromethyl, C₁₋₄alkyl, hydroxy,hydroxycarbonyl, or C₁₋₄alkyloxycarbonyl; R³ is a bivalent radical offormula

 wherein said (b-1)[, (b-2), or (b-3)] optionally can be substitutedwith one, two or three substituents each independently selected fromhalo, hydroxy, C₁₋₆alkyl, C₁₋₆alkyloxy, nitro, amino, cyano,trifluoromethyl, phenyl, or phenyl substituted with one or twosubsitutents each independently selected from halo, hydroxy, cyano,C₁₋₆alkyl, C₁₋₆alkyloxy, nitro, cyano, and trifluoromethyl.
 2. Acompound according to claim 1 wherein R² is (a-2).
 3. A compoundaccording to claim 1 wherein R¹ is C₁₋₆alkylcarbonyl,aminoC₁₋₆alkylcarbonyl or an amino acid.
 4. A pharmaceutical compositioncomprising a pharmaceutically acceptable carrier and a therapeuticallyactive amount of a compound as claimed in claim
 1. 5. A process forpreparing a compound of formula (I) wherein a) an intermediate offormula (II) is reacted with an intermediate of formula (III) in areaction-inert solvent and, optionally in the presence of a suitablebase, thereby yielding compounds of formula (I-a), defined as compoundsof formula (I) wherein R^(1a) represents all R¹ substituents other thanC₁₋₄alkyl substituted with amino; or

b) an intermediate of formula (II) is reacted with an intermediate offormula (IV), thereby yielding a compound of formula (I-a);

wherein in the above reaction schemes the radicals R¹, R², R³, and theinteger n, are as defined in claim 1; c) or, compounds of formula (I)are converted into each other following art-known transformationreactions; or if desired; a compound of formula (I) is converted into anacid addition salt, or conversely, an acid addition salt of a compoundof formula (I) is converted into a free base form with alkali; and, ifdesired, preparing stereochemically isomeric forms thereof.
 6. A methodof treating eating disorders or obesity comprising administering aneffective amount of a compound of Formula I as claimed in claim 1.